Sensitive detection of point mutation using exponential strand displacement amplification-based surface enhanced Raman spectroscopy
文献类型：期刊论文
| 作者 | Huang Siqiang; Hu Juan; Zhu Guichi; Zhang Chunyang |
| 刊名 | BIOSENSORS & BIOELECTRONICS
 |
| 出版日期 | 2015 |
| 英文摘要 | Accurate identification of point mutation is particularly imperative in the field of biomedical research and clinical diagnosis. Here, we develop a sensitive and specific method for point mutation assay using exponential strand displacement amplification (SDA)-based surface enhanced Raman spectroscopy (SERS). In this method, a discriminating probe and a hairpin probe are designed to specifically recognize the sequence of human K-ras gene. In the presence of K-ras mutant target (C -> T), the 3'-terminal of discriminating probe and the 5'-terminal of hairpin probe can be ligated to form a SDA template. Subsequently, the 3'-terminal of hairpin probe can function as a primer to initiate the SDA reaction, producing a large amount of triggers. The resultant triggers can further hybridize with the discriminating probes to initiate new rounds of SDA reaction, leading to an exponential amplification reaction. With the addition of capture probe-modified gold nanoparticles (AuNPs) and the Rox-labeled reporter probes, the amplified triggers can be assembled on the surface of AuNPs through the formation of sandwich hybrids of capture probe-trigger-reporter probe, generating a strong Raman signal. While in the presence of K-ras wild-type target (C), neither ligation nor SDA reaction can be initiated and no Raman signal is observed. The proposed method exhibits high sensitivity with a detection limit of 1.4 pM and can accurately discriminate as low as 1% variant frequency from the mixture of mutant target and wild-type target. Importantly, this method can be further applied to analyze the mutant target in the spiked HEK293T cell lysate, holding great potential for genetic analysis and disease prognosis. (C) 2014 Elsevier B.V. All rights reserved |
| 收录类别 | SCI |
| 原文出处 | http://www.ncbi.nlm.nih.gov/pubmed/25461157 |
| 语种 | 英语 |
| 源URL | [http://ir.siat.ac.cn:8080/handle/172644/7365]  |
| 专题 | 深圳先进技术研究院_医药所 |
| 作者单位 | BIOSENSORS & BIOELECTRONICS |
| 推荐引用方式 GB/T 7714 | Huang Siqiang,Hu Juan,Zhu Guichi,et al. Sensitive detection of point mutation using exponential strand displacement amplification-based surface enhanced Raman spectroscopy[J]. BIOSENSORS & BIOELECTRONICS,2015. |
| APA | Huang Siqiang,Hu Juan,Zhu Guichi,&Zhang Chunyang.(2015).Sensitive detection of point mutation using exponential strand displacement amplification-based surface enhanced Raman spectroscopy.BIOSENSORS & BIOELECTRONICS. |
| MLA | Huang Siqiang,et al."Sensitive detection of point mutation using exponential strand displacement amplification-based surface enhanced Raman spectroscopy".BIOSENSORS & BIOELECTRONICS (2015). |
入库方式： OAI收割
来源：深圳先进技术研究院
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